1. Evolution of a Behavioral Shift Mediated by Superficial Neuromasts Helps Cavefish Find Food in Darkness 
Masato Yoshizawa, Špela Gorički, Daphne Soares, William R. Jeffery 
Current Biology 
Volume 20, Issue 18, Pages (September 2010)
DOI: /j.cub
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2. Figure 1 Quantification of Vibration Attraction Behavior
(A) Diagram showing the assay system used to record vibration attraction behavior (VAB) in darkness. An infrared light illuminates the cylinder-shaped assay chamber from below. Swimming is recorded with a charge-coupled device infrared camera positioned above the assay chamber. A glass rod is inserted into the assay chamber to produce vibration stimuli.
(B and C) Path (purple lines) of swimming in Pachón cavefish (B) and surface fish (C) during the 3 min assay period. Dotted lines indicate 2 cm diameter quantification area surrounding the grass rod (dark spot in the center of the chamber). Arrows indicate the starting positions of the fish.
(D–F) VAB quantified in cavefish (orange lines and points) and surface fish (black lines and points) as number of approaches (NOA, D), duration (E), and latency (F) in the absence of a rod (left), the presence of a stationary rod (middle), and the presence of a vibrating rod (50 Hz, right). Values are means ± standard error of the mean (SEM). Cavefish: n = 40; surface fish: n = 44. Orange, black, and blue asterisks indicate significant differences among cavefish, among surface fish, and between cavefish and surface fish, respectively (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001). See also Movie S1, Figure S1, and Tables S1 and S2.
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3. Figure 2
Significance of VAB in Feeding Determined by Competitive Prey-Capture Experiments
(A) VAB levels in surface fish and Pachón cavefish measured as NOA. Surface fish: n = 54, gray area; cavefish: n = 52, orange area. Vertical dashed line represents NOA cutoff value for classifying fish with (above 4) and without (below 4) VAB with a stimulus of 50 Hz.
(Ba–Bc) Prey-capture competition assays. Bars show the proportion of strikes at prey between pairs of surface fish (gray bars) and cavefish (orange bars) with or without VAB during a 1 min assay period in darkness (Ba–Bc, left bars) and in light (Ba–Bc, right bars). A total of eight pairs of cavefish versus surface fish (Ba), five pairs of surface fish with and without VAB (Bb), and ten pairs of cavefish with and without VAB (Bc) in the dark and light are shown. Values are mean ratio of strikes ± 95% confidence intervals of the mean. ∗p < 0.05, ∗∗p < See also Table S1.
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4. Figure 3 VAB Is Controlled by Superficial Neuromasts
(A) The relationship between VAB and vibration frequencies (Hz) in Pachón cavefish and surface fish. Values are mean NOA ± SEM. Cavefish: n = 40 at no rod and 0 Hz; n = 35 at 5 Hz, 10 Hz, 20 Hz, 100 Hz, and 200 Hz; n = 19 at 35 Hz and 75 Hz; n = 51 at 50 Hz; n = 8 at 300 Hz; n = 27 at 500 Hz. Asterisks denote frequencies at which cavefish VAB is significantly above the 0 Hz condition at p < 0.05 (see Table S1). Surface fish: n = 44 at no rod and 0 Hz; n = 38 at 5 Hz, 10 Hz, 20 Hz, and 100 Hz; n = 19 at 35 Hz and 75 Hz; n = 54 at 50 Hz; n = 35 at 200 Hz; n = 8 at 300 Hz; n = 27 at 500 Hz. Open horizontal boxes indicate the best sensing range of superficial neuromasts (SN), canal neuromasts (CN), and inner ear (right) in Astyanax [18, 20]. See also Figures S2 and S3.
(B and C) DASPEI-stained Pachón cavefish (B) and surface fish (C) before and after SN ablation. Scale bar is 1.0 mm; magnification is the same in (B) and (C).
(D) VAB in cavefish and surface fish before SN ablation and 2 days after bilateral SN ablation. Values are means ± SEM. ∗p < 0.05, ∗∗p < Cavefish: n = 16 for SO-3, n = 10 for small region of the trunk, and n = 11 for large region of the trunk. Surface fish: n = 13 for SO-3 and n = 6 for small region of the trunk. A vibrating rod at 35 Hz was used. See also Table S1 and Figure S4.
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5. Figure 4
The Relationship between VAB and SN Number and Size in Pachón Cavefish, Surface Fish, and the F1 Progeny of a Surface Fish × Cavefish Cross
(A and B) Scatter plots showing the relationship between the SN number (A) or diameter (B) and the square root of NOA. Both SN number and diameter are significantly correlated with VAB (see Table S1). Cavefish: n = 20; surface fish: n = 19; F1 hybrids: n = 23. Linear regression line is in red. A vibrating rod at 35 Hz was used.
(C–E) DASPEI staining of neuromasts in the SO-3 region of a cavefish (C), an F1 hybrid (D), and a surface fish (E). Scale bar in (E) is 1.0 mm; magnification is the same in (C)–(E). Insets in (C)–(E) show the size of a typical SN from the SO-3 region of fish in (C)–(E). Scale bar in inset (E) is 50 μm; magnification is the same in each inset.
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